The glycerolipid/ free fatty acid (GL/FFA) cycle is a key metabolic pathway that links glucose and fatty acid metabolism and it consists of lipogenesis and lipolysis. GL/FFA cycling, especially in its lipolysis arm, generates various lipid signaling molecules to regulate insulin secretion in pancreatic ß-cells and non-shivering thermogenesis in adipocytes. Currently, the lipolysis-derived lipid signals involved in this process are uncertain.
Triglyceride hydrolysis in mammalian cells is accomplished by the sequential actions of adipose triglyceride lipase to produce diacylglycerol, by hormone sensitive lipase to produce monoacylglycerol (MAG) and by MAG lipase (MAGL) that releases free fatty acid and glycerol. Our work shows that in pancreatic ß-cell, the classical MAGL is poorly expressed and that MAG hydrolysis is mainly conducted by the newly identified α/β-Hydrolase Domain-6 (ABHD6). Inhibition of ABHD6 by its specific inhibitor WWL70, leads to long-chain saturated 1-MAG accumulation inside the cells, accompanied by enhanced glucose-stimulated insulin secretion (GSIS). Decreasing the MAG levels by overexpression of ABHD6 in the ß-cell line INS832/13 reduces GSIS, while increasing MAG levels by ABHD6 knockdown enhances GSIS. Acute exposure of INS832/13 cells to various MAG species dose-dependently stimulates insulin secretion and restores GSIS suppressed by the pan-lipase inhibitor orlistat. Also, various biochemical and pharmacological experiments show that saturated 1-MAG levels species rather than unsaturated or 2-MAG species best correlate with insulin secretion. Furthermore, whole-body and β-cell-specific ABHD6-KO mice exhibit enhanced GSIS in vivo, and their isolated islets show elevated MAG production and GSIS. Inhibition of ABHD6 in low dose streptozotocin diabetic mice restores GSIS and improves glucose tolerance. Results further show that ABHD6-accessible MAGs not only enhance GSIS, but also potentiate fatty acid and non-fuel-induced insulin secretion without alteration in glucose oxidation and utilization as well as fatty acid oxidation. We have identified that MAG binds and activates the vesicle priming protein Munc13-1, thereby inducing insulin exocytosis. Based on all these observations, we propose that lipolysis-derived saturated 1-MAG acts as a metabolic coupling factor to regulate insulin secretion and ABHD6 is a negative modulator of insulin secretion.
Besides its role in ß-cells, ABHD6 is also highly expressed in adipocytes and its level is increased with obesity. Mice globally lacking ABHD6 on high fat diet (HFD) show modestly reduced food intake, decreased body weight gain, insulinemia and fasting glycemia and improved glucose tolerance and insulin sensitivity and enhanced locomotor activity. In addition, ABHD6-KO mice display increased energy expenditure and cold-induced thermogenesis. In accordance with this, these mice show elevated UCP1 level in white and brown adipocytes, indicating browning of white adipocytes. The browning phenotype is reproduced in the mice either chronically treated with the ABHD6 inhibitor WWL70 or an antisense oligonucleotides targeting ABHD6. White and brown adipose tissues isolated from whole body ABHD6 KO mice show greatly elevated levels of 1-MAG, but not 2-MAG. Increasing MAG levels by either exogenous administration of 1-MAG or ABHD6 inhibition or genetic deletion induces browning of white adipocytes in a cell-autonomous manner. Further evidence indicates that 1-MAGs can transactivate PPARα and PPARγ and the browning effect induced by WWL70 or exogenous MAG is abolished by PPARα and PPARγ antagonists. In vivo administration of the PPARα antagonist GW6471 to ABHD6 KO mice partially reversed the ABHD6-KO effects on body weight gain, and abolishes the enhanced thermogenesis, white adipose browning and fatty acid oxidation in brown adipose tissue. All these observations indicate that ABHD6 regulates not only insulin and glucose homeostasis but also energy homeostasis and adipose tissue function.
Thus, ABHD6-accessible 1-MAG not only acts as a metabolic coupling factor to regulate fuel and non-fuel induced insulin secretion by activating Munc13-1 in beta cells, but also regulates glucose, insulin and energy homeostasis. The latter effects are mediated at least in part via browning of white adipocytes and enhanced brown fat function through the activation of PPARα and PPARγ. Collectively these findings suggest that ABHD6 is a promising target for developing therapeutics against obesity, type 2 diabetes and metabolic syndrome.